The present invention relates to a hair-growing agent comprising a phosphatidic acid as an active ingredient.
A known example of a cosmetic or drug relating to hair which comprises a phosphatidic acid is a hair-nourishing agent comprising a phosphatidic acid having straight-chain fatty acid residues having an odd number of carbon atoms, that is, a phosphatidic acid represented by formula (II): 
(wherein R3 and R4 each represents an aliphatic hydrocarbon group; and at least one of R3 and R4 is a straight-chain aliphatic hydrocarbon group having an even number of carbon atoms) (Japanese Published Examined Patent Application No. 41363/88). Japanese Published Unexamined Patent Application No. 7205/86 describes a cell activator comprising a phosphatidic acid having two branched-chain fatty acid residues as an active ingredient. However, there has not been known a hair-growing agent comprising a phosphatidic acid having at least one ether group. Japanese Published Unexamined Patent Application No. 15809/86 describes a cell activator comprising a fatty acid having an odd number of carbon atoms, and biotin or vitamin B12 as active ingredients.
It is known that a phosphatidic acid is used as a vehicle for liposome preparation added to minoxidil which is a hair growth-activating component (U.S. Pat. No. 5,030,442).
A mixture of some phospholipids including a phosphatidic acid is known to have hair loss-inhibiting activity (WO 97/09989). Also known is a hair-growing agent comprising a phospholipid mixture (DE 3222016, DE 4113346).
WO 96/00561 describes a hair-growing agent comprising proanthocyanidin. It is known that tocopherol [xe2x80x9cKe no Igakuxe2x80x9d (Medical Science of Hair), p. 283, Bunkodo (1987); Hair Science, p. 80, Japan Hair Science Association (1986)], pantothenic acid and biotin [Fragrance Journal, p. 95-103, Fragrance Journal (1989)], and a protein kinase C-specific inhibitor [Skin Pharmacology and Applied Skin Physiology, 13, 133-142 (2000)] each have hair-growing activity.
Japanese Published Unexamined Patent Application No. 263711/99 describes the hair-growing activity of 1-O-hexadecyl-2-O-methylglycerol.
However, there is no report on a hair-growing agent comprising, as active ingredients, a phosphatidic acid represented by formula (I) shown hereinbelow, and one or more members selected from the group consisting of proanthocyanidin, tocopherol, derivatives of tocopherol, pantothenic acid, derivatives of pantothenic acid, protein kinase C-specific inhibitors or pharmaceutically acceptable salts thereof and biotin; a hair-growing agent comprising, as active ingredients, a phosphatidic acid, and one or more members selected from the group consisting of proanthocyanidin, tocopherol, derivatives of tocopherol, pantothenic acid, derivatives of pantothenic acid and protein kinase C-specific inhibitors or pharmaceutically acceptable salts thereof; or a hair-growing agent comprising, as active ingredients, a phosphatidic acid and biotin, and one or more members selected from the group consisting of proanthocyanidin, tocopherol, derivatives of tocopherol, pantothenic acid, derivatives of pantothenic acid and protein kinase C-specific inhibitors or pharmaceutically acceptable salts thereof.
The present invention provides a hair-growing agent comprising a phosphatidic acid as an active ingredient, which has an excellent hair-growing effect; a hair-growing agent comprising, as active ingredients, a phosphatidic acid, and one or more members selected from the group consisting of proanthocyanidin, tocopherol, derivatives of tocopherol, pantothenic acid, derivatives of pantothenic acid, protein kinase C-specific inhibitors or pharmaceutically acceptable salts thereof and biotin; and the like.
The present invention relates to the following subject matters.
(1) A hair-growing agent comprising, as an active ingredient, a phosphatidic acid represented by formula (I): 
xe2x80x83(wherein R1 represents alkyl, alkenyl, alkanoyl or alkenoyl; and when R1 represents alkyl or alkenyl, R2 represents alkyl, alkenyl, alkanoyl or alkenoyl, and when R1 represents alkanoyl or alkenoyl, R2 represents alkyl or alkenyl).
(2) The hair-growing agent according to the above (1), wherein R1 is alkyl or alkenyl, and R2 is methyl.
(3) The hair-growing agent according to the above (1), wherein R1 is alkanoyl or alkenoyl, and R2 is methyl.
(4) The hair-growing agent according to the above (1), wherein R1 is alkyl or alkenyl, and R2 is acetyl.
(5) The hair-growing agent according to the above (1), wherein R1 is octadecenoyl or hexadecyl, and R2 is methyl.
(6) The hair-growing agent according to the above (1), wherein R1 is hexadecyl and R2 is acetyl.
(7) A hair-growing agent comprising, as active ingredients, the phosphatidic acid according to any of the above (1) to (6), and one or more members selected from the group consisting of proanthocyanidin, tocopherol, derivatives of tocopherol, pantothenic acid, derivatives of pantothenic acid, protein kinase C-specific inhibitors or pharmaceutically acceptable salts thereof and biotin.
(8) The hair-growing agent according to any of the above (1) to (6), further comprising proanthocyanidin.
(9) The hair-growing agent according to the above (8), wherein the proanthocyanidin is one or more members selected from the group consisting of procyanidin B-1, procyanidin B-2, procyanidin B-3, procyanidin C-1 and procyanidin C-2.
(10) The hair growing agent according to any of the above (1) to (6), (8) and (9), further comprising tocopherol or a derivative of tocopherol.
(11) The hair-growing agent according to the above (10), wherein the tocopherol or the derivative of tocopherol is one or more members selected from the group consisting of dl-xcex1-tocopherol, d-xcex1-tocopherol, dl-xcex1-tocopherol acetate, d-xcex1-tocopherol acetate and dl-xcex1-tocopherol nicotinate.
(12) The hair-growing agent according to any of the above (1) to (6) and (8) to (11), further comprising pantothenic acid or a derivative of pantothenic acid.
(13) The hair-growing agent according to the above (12), wherein the pantothenic acid or the derivative of pantothenic acid is one or more members selected from the group consisting of calcium pantothenate, sodium pantothenate, D-pantothenyl alcohol, DL-pantothenyl alcohol and pantothenyl ethyl ether.
(14) The hair-growing agent according to any of the above (1) to (6) and (8) to (13), further comprising a protein kinase C-specific inhibitor or a pharmaceutically acceptable salt thereof.
(15) The hair-growing agent according to the above (14), wherein the protein kinase C-specific inhibitor is one or more members selected from the group consisting of calphostin C, hexadecylphosphocholine, palmitoyl-DL-carnitine and polymyxin B.
(16) The hair-growing agent according to any of the above (1) to (6) and (8) to (15), further comprising biotin.
(17) The hair-growing agent according to any of the above (1) to (16), which does not substantially comprise minoxidil.
(18) A hair-growing agent comprising, as active ingredients, a phosphatidic acid, and one or more members selected from the group consisting of proanthocyanidin, tocopherol, derivatives of tocopherol, pantothenic acid, derivatives of pantothenic acid and protein kinase C-specific inhibitors or pharmaceutically acceptable salts thereof.
(19) A hair-growing agent comprising, as active ingredients, a phosphatidic acid and biotin, and one or more members selected from the group consisting of proanthocyanidin, tocopherol, derivatives of tocopherol, pantothenic acid, derivatives of pantothenic acid and protein kinase C-specific inhibitors or pharmaceutically acceptable salts thereof.
(20) A method for stimulating hair growth in a mammal, which comprises applying to the skin of said mammal a phosphatidic acid represented by formula (I): 
xe2x80x83(wherein R1 represents alkyl, alkenyl, alkanoyl or alkenoyl; and when R1 represents alkyl or alkenyl, R2 represents alkyl, alkenyl, alkanoyl or alkenoyl, and when R1 represents alkanoyl or alkenoyl, R2 represents alkyl or alkenyl).
(21) A method for stimulating hair growth in a mammal, which comprises applying to the skin of said mammal the phosphatidic acid according to the above (20), and one or more members selected from the group consisting of proanthocyanidin, tocopherol, derivatives of tocopherol, pantothenic acid, derivatives of pantothenic acid, protein kinase C-specific inhibitors or pharmaceutically acceptable salts thereof and biotin.
In the definitions of each group in formula (I), the alkyl includes alkyl groups having 1 to 24, preferably 7 to 19 carbon atoms, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, heneicosyl, docosyl, tricosyl and tetracosyl. The alkenyl includes alkenyl groups having 2 to 24, preferably 7 to 19 carbon atoms, such as vinyl, allyl, butenyl, 2-pentenyl, 4-pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodecenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, nonadecenyl, eicosenyl, heneicosenyl, docosenyl, tricosenyl and tetracosenyl. The number and position of unsaturated bonds in the alkenyl are not specifically restricted. The alkanoyl includes alkanoyl groups having 2 to 24, preferably 7 to 20 carbon atoms, such as acetyl, propionyl, butyryl, valeryl, hexanoyl, heptanoyl, octanoyl, nonanoyl, decanoyl, undecanoyl, dodecanoyl, tridecanoyl, tetradecanoyl, pentadecanoyl, hexadecanoyl, heptadecanoyl, octadecanoyl, nonadecanoyl, eicosanoyl, heneicosanoyl, docosanoyl, tricosanoyl and tetracosanoyl. The alkenoyl includes alkenoyl groups having 3 to 24, preferably 7 to 20 carbon atoms, such as propenoyl, butenoyl, 2-pentenoyl, 4-pentenoyl, hexenoyl, heptenoyl, octenoyl, nonenoyl, decenoyl, undecenoyl, dodecenoyl, tridecenoyl, tetradecenoyl, pentadecenoyl, hexadecenoyl, heptadecenoyl, cis-9-octadecenoyl, cis-9-cis-12-octadecadienoyl, 9,12,15-octadecatrienoyl, nonadecenoyl, 5,8,11,14-eicosatetraenoyl, heneicosenoyl, docosenoyl, tricosenoyl and tetracosenoyl. The number and position of unsaturated bonds in the alkenoyl are not specifically restricted.
The phosphatidic acids to be used in the present invention can be obtained mainly by chemical synthesis, that is, by introducing a fatty acid or alcohol to the 1-position of glycerin and then introducing the same or another fatty acid or alcohol to the 2-position of glycerin, followed by phosphorylation of glycerin at the 3-position. In each step of the synthesis, a protecting group may be introduced as may be required. For example, the desired phosphatidic acids can be obtained in the following manner. To a mixture of 1 to 5 equivalents of phosphorus oxychloride, 1 to 5 equivalents of triethylamine and hexane is added dropwise 1 to 5 equivalents of a derivative of glycerol in an inert solvent. The resulting mixture is stirred at a temperature below the boiling point of the solvent, preferably under ice cooling, for 10 to 20 hours, and purified water is added thereto. After the resulting mixture is stirred at room temperature for one hour, the organic layer is washed and concentrated under reduced pressure, whereby the desired phosphatidic acid is obtained. The inert solvent includes chloroform and hexane. The desired phosphatidic acids can also be obtained by enzymatic hydrolysis of a phospholipid derivative containing a fatty acid residue or an alcohol residue. For example, when a derivative of phosphatidyl choline is employed as a starting material, the desired phosphatidic acid can be obtained by hydrolyzing the phosphate bond with choline using an enzyme such as phospholipase D [Harumi Okuyama, Biochemistry of Lipids (Lectures on Biochemical Experiments 3), The Japanese Biochemical Society, p. 289, Tokyo Kagaku Dojin (1974)]. However, the methodology for the production of phosphatidic acids is not limited to the above-mentioned methods, and the above phosphatidic acids obtained by any methods may be used in the present invention.
The proanthocyanidin used in the present invention is a polymer composed of flavan-7-ol derivatives represented by formula (III): 
(wherein R5 and R6, which may be the same or different, each represents a hydrogen atom or a hydroxyl group) or the like as constitutive units.
Examples of the flavan-7-ol derivatives include catechin, epicatechin, gallocatechin, epigallocatechin, afzelechin, epiafzelechin and any optical isomers thereof. Proanthocyanidin composed of epicatechin or catechin as a constitutive unit is preferably used in the present invention.
The bonding mode of the flavan-7-ol derivatives represented by formula (III) may be any mode. An example of a dimer composed of two flavan-7-ol derivatives is the one which has a bonding mode represented by formula (IV): 
(wherein R7 and R7a have the same significance as the above R5, and R8 and R8a have the same significance as the above R6). In trimers and higher polymers, the flavan-7-ol derivatives may be bonded in the same or different bonding modes.
The proanthocyanidin to be used in the present invention may be any dimer or higher polymer of flavan-7-ol derivatives, and is preferably a 2- to 10-mer, more preferably a 2- to 5-mer, further preferably a dimer or trimer. Examples of the dimers of flavan-7-ol derivatives include epicatechin-catechin co-dimers such as epicatechin-(4xcex2xe2x86x928)-catechin, epicatechin dimers such as epicatechin-(4xcex2xe2x86x928)-epicatechin, and catechin dimers such as catechin-(4xcex1xe2x86x928)-catechin. Examples of the trimers of flavan-7-ol derivatives include epicatechin trimers such as epicatechin-(4xcex2xe2x86x928)-epicatechin-(4xcex2xe2x86x928)-epicatechin, catechin trimers such as catechin-(4xcex1xe2x86x928)-catechin-(4xcex1xe2x86x928)-catechin, and epicatechin-catechin co-trimers such as epicatechin-(4xcex2xe2x86x928)-epicatechin-(4xcex2xe2x86x928)-catechin.
The proanthocyanidin to be used in the present invention also includes compounds wherein gallic acid or a sugar such as glucose or rhamnose is attached to the above proanthocyanidin.
Proanthocyanidin can be obtained by extraction and purification from various plants such as grape, apple, barley, Japanese persimmon, coconut, cacao, pine, azuki bean and peanut belonging to the genera Vitis, Malus, Hordeum, Diospyros, Cocos, Theobroma, Pinus, Phaseolus, Arachis and the like, or by chemical synthesis.
For instance, proanthocyanidin can be extracted and purified from plants according to the following known method.
Fruits, seeds, leaves, stalks, roots, rootstocks, etc. of the plants as starting materials are harvested at a suitable stage and used, as such or usually after being subjected to drying such as air drying, as materials for extraction. Extraction of proanthocyanidin from dry plants can be carried out in a manner similar to known methods [Chemical and Pharmaceutical Bulletin, 3, 3218 (1990); ibid., 40, 889-898 (1992)].
That is, the materials are ground or cut into fine pieces, followed by extraction with a solvent. Suitable solvents for extraction include hydrophilic or lipophilic solvents such as water, alcohols (e.g. ethanol, methanol and isopropyl alcohol), ketones (e.g. acetone and methyl ethyl ketone) and esters (e.g. methyl acetate and ethyl acetate), which can be used alone or as a mixture. The temperature for extraction is usually 0 to 100xc2x0 C., preferably 5 to 50xc2x0 C. The time for extraction is about one hour to 10 days, and the amount of the solvent is usually 1 to 30 times by weight, preferably 5 to 10 times by weight based on the dry material. Extraction is carried out by stirring or by dipping followed by standing, and is repeated twice or 3 times, as may be required.
The crude extract obtained in the above manner is filtered or centrifuged to remove the insoluble residue. Purification of proanthocyanidin from the thus treated extract, or from juice or sap of the plants can be carried out by any known purification methods. It is preferred to employ the two-phase solvent partitioning method, column chromatography, preparative high-performance liquid chromatography, etc. alone or in combination. The two-phase solvent partitioning methods include, for example, a method in which oil-soluble components and pigments are removed from the above extract by extraction with n-hexane, petroleum ether, etc., and a method in which proanthocyanidin is collected from the extract into the solvent phase by partition between a solvent such as n-butanol or methyl ethyl ketone and water. Column chromatography includes a method using normal phase silica gel, a method using reversed phase silica gel, adsorption column chromatography using as a carrier Diaion HP-20, Sepabeads SP-207 or the like, and gel filtration using as a carrier Sephadex LH-20 or the like. They are employed alone or in combination, if necessary repeatedly. Preparative high-performance liquid chromatography includes a method using a reversed phase column packed with octadecyl silica or the like, and a method using a normal phase column packed with silica gel or the like.
Proanthocyanidin can be purified by removing water-soluble ionic substances such as salts, nonionic substances such as saccharides and polysaccharides, oils, pigments, etc. from the above extract according to the above purification methods.
Grape-derived proanthocyanidin can be extracted and purified according to the method described in Acta Dermato Venereologica, 78, 428-432 (1998) or a similar method. Procyanidin B-1 [epicatechin-(4xcex2xe2x86x928)-catechin], procyanidin B-2 [epicatechin-(4xcex2xe2x86x928)-epicatechin], procyanidin B-3 [catechin-(4xcex1xe2x86x928)-catechin], procyanidin C-1 [epicatechin-(4xcex2xe2x86x928)-epicatechin-(4xcex2xe2x86x928)-epicatechin] and procyanidin C-2 [catechin-(4xcex1xe2x86x928)-catechin-(4xcex1xe2x86x928)-catechin] can be extracted and purified according to the method described in The Journal of Investigative Dermatology, 112, 310-316 (1999) or a similar method.
Production of proanthocyanidin by chemical synthesis can be carried out according to the method described in Journal of Chemical Society, Perkin Transaction I, 1535-1543 (1983) in which a process of producing dimers of epicatechin or catechin is described, the method described in Phytochemistry, 25, 1209-1215 (1986) or similar methods.
When proanthocyanidin is used as an active ingredient in the present invention, one or more kinds of proanthocyanidin may be used alone or as a mixture. It is preferred to use one or more members selected from the group consisting of grape-seed-derived proanthocyanidin, apple-derived proanthocyanidin, barley-derived proanthocyanidin, pine-derived proanthocyanidin, purified procyanidin oligomers, procyanidin B-1, procyanidin B-2, procyanidin B-3, procyanidin C-1 and procyanidin C-2. Specifically it is preferred to use one or more members selected from the group consisting of procyanidin B-1, procyanidin B-2, procyanidin B-3, procyanidin C-1 and procyanidin C-2.
The tocopherol and derivatives of tocopherol to be used in the present invention include any natural and synthetic ones that are commercially available, and derivatives such as acetic acid esters and nicotinic acid esters. Examples thereof include dl-xcex1-tocopherol, d-xcex1-tocopherol, dl-xcex1-tocopherol acetate, d-xcex1-tocopherol acetate and dl-xcex1-tocopherol nicotinate.
The pantothenic acid and derivatives of pantothenic acid to be used in the present invention include any natural and synthetic ones that are commercially available and any other ones. Examples thereof include calcium pantothenate, sodium pantothenate, D-pantothenyl alcohol, DL-pantothenyl alcohol and pantothenyl ethyl ether.
As the protein kinase C-specific inhibitor in the present invention, any inhibitor that specifically inhibits protein kinase C can be used. It is preferred to use protein kinase inhibitors of which the ratio of the 50% protein kinase A (PKA) inhibition constant (hereinafter referred to as PKA-IC50) to the 50% protein kinase C (PKC) inhibition constant (hereinafter referred to as PKC-IC50) (the ratio is hereinafter referred to as PKA-IC50/PKC-IC50) is 3 or more, preferably 3 to 109, more preferably 10 to 109, when PKC-inhibiting activity and PKA-inhibiting activity are measured by the following methods for measuring PKC-inhibiting activity and PKA-inhibiting activity. Examples thereof are one or more members selected from the group consisting of calphostin C, hexadecylphosphocholine, palmitoyl-DL-carnitine, polymyxin B and pharmaceutically acceptable salts thereof.
Examples of the pharmaceutically acceptable salts are hydrochlorides, hydrobromides, sulfates, nitrates, formates, acetates, benzoates, maleates, fumarates, succinates, tartrates, citrates, oxalates, methanesulfonates, toluenesulfonates, aspartates and glutamates.
The methods for measuring PKC-inhibiting activity and PKA-inhibiting activity are described below.
(1) Method for Measuring PKC-inhibiting Activity
Measurement of PKC-inhibiting activity can be carried out in a manner similar to the method of Kikkawa, et al. [Journal of Biological Chemistry, 257, 13341 (1982)].
To 250 xcexcl of an aqueous solution comprising 2.5 xcexcmol of magnesium acetate, 50 xcexcg of histone Type IIIS (Sigma Chemical Co., Ltd.), 20 xcexcg of phosphatidylserine, 0.8 xcexcg of diolein, 25 nmol of calcium chloride, 5 xcexcg of a crude enzyme (partially purified from rat brain by the method of Kikkawa, et al.) and 5 xcexcmol of Tris-HCl buffer (pH 7.5) is added the above aqueous solution containing a test compound (10 xcexcl), followed by incubation at 30xc2x0 C. for 3 minutes. After the incubation, 1.25 nmol of [xcex3-32P]ATP (5-10xc3x97103 cpm/nmol) is added thereto, followed by phosphorylation reaction at 30xc2x0 C. for 3 minutes. The reaction is terminated by addition of 25% trichloroacetic acid and the reaction mixture is filtered through a cellulose acetate membrane (pore size: 0.45 xcexcm, Toyo Filter Co., Ltd.) After the membrane is washed four times with 5% trichloroacetic acid, the radioactivity remaining on the membrane is measured as a test compound value. Separately, the above procedure is carried out in the same manner without addition of the test compound and the radioactivity is measured as a control value.
The molar concentration of the test compound giving a test compound value which is 50% of the control value is regarded as the 50% PKC inhibition constant (PKC-IC50).
(2) Method for Measuring PKA-inhibiting Activity
Measurement of PKA-inhibiting activity can be carried out in a manner similar to the method of Kuo, et al. [Biochemistry, 64, 1349 (1969)].
To 250 xcexcl of an aqueous solution comprising 5 xcexcmol of Tris-HCl buffer (pH 6.8), 2.5 xcexcmol of magnesium acetate, 100 xcexcg of histone Type IIS (Sigma Chemical Co., Ltd.), 0.25 nmol of c-AMP and 200 xcexcg of a crude enzyme (partially purified from calf heart by the method of Kuo, et al.) is added the above aqueous solution containing a test compound (10 xcexcl), followed by incubation at 30xc2x0 C. for 3 minutes. After the incubation, 1.25 nmol of [xcex3-32P]ATP (5-10xc3x97103 cpm/nmol) is added thereto, followed by phosphorylation reaction at 30xc2x0 C. for 3 minutes. The reaction is terminated by addition of 25% trichloroacetic acid and the reaction mixture is filtered through a cellulose acetate membrane (pore size: 0.45 xcexcm, Toyo Filter Co., Ltd.) After the membrane is washed four times with 5% trichloroacetic acid, the radioactivity remaining on the membrane is measured as a test compound value. Separately, the above procedure is carried out in the same manner without addition of the test compound and the radioactivity is measured as a control value.
The molar concentration of the test compound giving a test compound value which is 50% of the control value is regarded as the 50% PKA inhibition constant (PKA-IC50).
As the biotin, any natural or synthetic biotin that is commercially available can be used. Suitable examples thereof include D-biotin.
The hair-growing agent of the present invention may be in any preparation form so long as it can contain a phosphatidic acid represented by formula (I); the phosphatidic acid, and one or more members selected from the group consisting of proanthocyanidin, tocopherol, derivatives of tocopherol, pantothenic acid, derivatives of pantothenic acid, protein kinase C-specific inhibitors or pharmaceutically acceptable salts thereof and biotin; a phosphatidic acid, and one or more members selected from the group consisting of proanthocyanidin, tocopherol, derivatives of tocopherol, pantothenic acid, derivatives of pantothenic acid and protein kinase C-specific inhibitors or pharmaceutically acceptable salts thereof; or a phosphatidic acid and biotin, and one or more members selected from the group consisting of proanthocyanidin, tocopherol, derivatives of tocopherol, pantothenic acid, derivatives of pantothenic acid and protein kinase C-specific inhibitors or pharmaceutically acceptable salts thereof. For example, it can be used in the form of a liquid or solid hair-growing preparation containing a suitable pharmaceutical vehicle.
Examples of the liquid or solid hair-growing preparations include liquid preparations such as hair liquid, hair tonic and hair lotion, and solid preparations such as ointment and hair cream. These preparations can be produced by adding to a suitable vehicle a phosphatidic acid represented by formula (I); the phosphatidic acid, and one or more members selected from the group consisting of proanthocyanidin, tocopherol, derivatives of tocopherol, pantothenic acid, derivatives of pantothenic acid, protein kinase C-specific inhibitors or pharmaceutically acceptable salts thereof and biotin; a phosphatidic acid, and one or more members selected from the group consisting of proanthocyanidin, tocopherol, derivatives of tocopherol, pantothenic acid, derivatives of pantothenic acid and protein kinase C-specific inhibitors or pharmaceutically acceptable salts thereof; or a phosphatidic acid and biotin, and one or more members selected from the group consisting of proanthocyanidin, tocopherol, derivatives of tocopherol, pantothenic acid, derivatives of pantothenic acid and protein kinase C-specific inhibitors or pharmaceutically acceptable salts thereof, according to a conventional method.
The content of a phosphatidic acid in the hair-growing agent of the present invention widely varies depending upon the kind of the phosphatidic acid and the percutaneous absorbability derived from physical properties, but it is usually, alone or as a mixture, 0.01 to 5.0 wt % (hereinafter referred to merely as %), preferably 0.01 to 3.0%, more preferably 0.1 to 1.0%. The content of proanthocyanidin varies depending upon the purification degree, but is usually 0.01 to 10.0%, preferably 0.1 to 5.0%, more preferably 0.3 to 2.0%. The content of tocopherol or a derivative of tocopherol is usually 0.01 to 2%, preferably 0.05 to 2%, more preferably 0.05 to 1%. The content of pantothenic acid or a derivative of pantothenic acid is usually 0.01 to 2%, preferably 0.05 to 1%, more preferably 0.1 to 0.5%. The content of a protein kinase C-specific inhibitor or a pharmaceutically acceptable salt thereof widely varies depending upon the inhibitory activity and the percutaneous absorbability derived from physical properties, but it is usually, alone or as a mixture, 0.00001 to 1%, preferably 0.0001 to 1%, more preferably 0.001 to 0.1%. The content of biotin is usually 0.0001 to 0.1%, preferably 0.001 to 0.1%, more preferably 0.001 to 0.05%.
Preferred vehicles for liquid preparations are those which are generally used in hair-growing agents such as purified water, ethyl alcohol and polyvalent alcohols. If necessary, additives may be added thereto.
Examples of the polyvalent alcohols are glycerol, 1,3-butylene glycol and propylene glycol.
Additives include surfactants, vitamins, anti-inflammatory agents, microbicides, hormones, crude drug extracts, tinctures, refrigerants, moisturizers, antioxidants, sequestering agents and perfumes.
Examples of the surfactants are polyoxyethylene (60) hardened castor oil, polyoxyethylene (8) oleyl ether, polyoxyethylene (10) oleyl ether, polyoxyethylene (10) monooleate, polyoxyethylene (30) glyceryl monostearate, sorbitan monostearate, polyoxyethylene (20) sorbitan monooleate, sucrose fatty acid esters, hexaglycerin monooleate, hexaglycerin monolaurate, polyoxyethylene reduced lanolin, polyoxyethylene (20) lanolin alcohol, polyoxyethylene (25) glyceryl pyroglutamate isostearate, and N-acetylglutamine isostearyl ester.
Examples of the vitamins are benzyl nicotinate, nicotinamide, pyridoxine hydrochloride and riboflavin.
Examples of the anti-inflammatory agents are dipotassium glycyrrhizinate, xcex2-glycyrrhetinic acid, allantoin, diphenhydramine hydrochloride, guaiazulene and l-menthol.
Examples of the microbicides are trichlorohydroxydiphenyl ether, hinokitiol, triclosan, chlorohexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, zinc pyrithione, benzalkonium chloride, photosensitizing dye No. 301 and sodium mononitroguaiacol.
Examples of the hormones are ethynylestradiol, estrone and estradiol.
Examples of the crude drug extracts are extract of Swertia japonica Makino, garlic extract, ginseng extract, aloe extract, cinchona extract, plant worm extract and saffron extract.
Examples of the tinctures are capsicum tincture, ginger tincture and cantharis tincture.
Examples of the refrigerants are capsicum tincture, l-menthol and camphor.
Examples of the moisturizers are L-pyrrolidonecarboxylic acid, sodium hyaluronate and chondroitin sulfate.
Examples of the antioxidants are butylhydroxyanisole, isopropyl gallate, propyl gallate and erythorbic acid.
Examples of the sequestering agents are ethylenediamine tetraacetate and salts thereof.
Examples of the perfumes are natural perfumes such as orange oil, lemon oil, bergamot oil, lime oil, lemongrass oil and lavender oil, and synthetic perfumes such as menthol, rose oxide, linalool, citral and linalyl acetate.
When the above liquid preparations are used as spray, they can be used in combination with combustible gas, incombustible gas, or the like. Examples of the combustible gas are LPG (a liquefied petroleum gas) and dimethyl ether, and examples of the incombustible gas are a nitrogen gas and a carbon dioxide gas.
Vehicles for solid preparations include Vaseline, solid paraffin, vegetable oils, mineral oils, lanolin, wax and macrogol. Further, the above additives, and lower alcohols such as ethyl alcohol and isopropyl alcohol may be added thereto, if necessary.
The dose of the hair-growing agent of the present invention varies depending upon the age or the body weight of a patient, the symptom of the disease, the therapeutic effect, the mode of administration, the time of treatment or the like. The agent is percutaneously administered in an amount of 0.1 to 250 mg, preferably 1 to 100 mg in terms of phosphatidic acid per adult once to several times per day.
Certain embodiments of the present invention are illustrated in the following examples.